SLAS Discovery最新文献

{"title":"Compound management in a virtual research organization: The cornerstone of a reliable MMV discovery engine","authors":"Anna Adam,&nbsp;Elodie Chenu,&nbsp;Dominique Besson","doi":"10.1016/j.slasd.2024.100168","DOIUrl":"10.1016/j.slasd.2024.100168","url":null,"abstract":"<div><p>Despite the efforts towards malaria eradication, latest estimates show that the number of malaria cases is still rising, and malaria continues to have a devastating impact on people's health and livelihoods particularly in populations located in sub-Saharan Africa ¹. As a Product Development Partnership (PDP), MMV Medicines for Malaria Venture (MMV) plays a crucial role by using public and philanthropic funds to engage the pharmaceutical industry and academic research institutions to discover, develop and deliver the new drugs needed to control and eradicate malaria. MMV Discovery, working with partners, has developed a robust pipeline of molecules and a reliable discovery engine able to support research projects from screening to candidate nomination, providing access to centers of expertise and evaluating the profile and potential of molecules. To efficiently support this malaria discovery effort, MMV and its partners have established a state-of-the-art compound management network, supporting all discovery activities. This network serves both discovery projects and open innovation initiatives, such as MMV Open, tailoring workflows to align with distinct project objectives. In addition to this, MMV has implemented reliable integrated logistic tools and interfaces. These tools enable the efficient management and tracking of individual not solubilized (dry) samples of project compounds, as well as dedicated, solubilized libraries of compounds designated for primary screens targeting malaria and other neglected diseases.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 5","pages":"Article 100168"},"PeriodicalIF":3.1,"publicationDate":"2024-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000303/pdfft?md5=938ea9a32dbd6548b3ddd7c3c0aacd0c&pid=1-s2.0-S2472555224000303-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141312508","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a live cell assay for the zinc transporter ZnT8","authors":"Lucia Azzollini ,&nbsp;Dolores Del Prete ,&nbsp;Gernot Wolf ,&nbsp;Christoph Klimek ,&nbsp;Mattia Saggioro ,&nbsp;Fernanda Ricci ,&nbsp;Eirini Christodoulaki ,&nbsp;Tabea Wiedmer ,&nbsp;Alvaro Ingles-Prieto ,&nbsp;Giulio Superti-Furga ,&nbsp;Lia Scarabottolo","doi":"10.1016/j.slasd.2024.100166","DOIUrl":"10.1016/j.slasd.2024.100166","url":null,"abstract":"<div><p>Zinc is an essential trace element that is involved in many biological processes and in cellular homeostasis. In pancreatic β-cells, zinc is crucial for the synthesis, processing, and secretion of insulin, which plays a key role in glucose homeostasis and which deficiency is the cause of diabetes. The accumulation of zinc in pancreatic cells is regulated by the solute carrier transporter SLC30A8 (or Zinc Transporter 8, ZnT8), which transports zinc from cytoplasm in intracellular vesicles. Allelic variants of SLC30A8 gene have been linked to diabetes. Given the physiological intracellular localization of SLC30A8 in pancreatic β-cells and the ubiquitous endogenous expression of other Zinc transporters in different cell lines that could be used as cellular model for SLC30A8 recombinant over-expression, it is challenging to develop a functional assay to measure SLC30A8 activity. To achieve this goal, we have firstly generated a HEK293 cell line stably overexpressing SLC30A8, where the over-expression favors the partial localization of SLC30A8 on the plasma membrane. Then, we used the combination of this cell model, commercial FluoZin-3 cell permeant zinc dye and live cell imaging approach to follow zinc flux across SLC30A8 over-expressed on plasma membrane, thus developing a novel functional imaging- based assay specific for SLC30A8. Our novel approach can be further explored and optimized, paving the way for future small molecule medium-throughput screening.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 5","pages":"Article 100166"},"PeriodicalIF":3.1,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000285/pdfft?md5=cc84c4d2988c7d8585e18309f5ebba58&pid=1-s2.0-S2472555224000285-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141289026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel thin plate spline methodology to model tissue surfaces and quantify tumor cell invasion in organ-on-chip models","authors":"Elizabeth Elton ,&nbsp;Carly Strelez ,&nbsp;Nolan Ung ,&nbsp;Rachel Perez ,&nbsp;Kimya Ghaffarian ,&nbsp;Danielle Hixon ,&nbsp;Naim Matasci ,&nbsp;Shannon M. Mumenthaler","doi":"10.1016/j.slasd.2024.100163","DOIUrl":"10.1016/j.slasd.2024.100163","url":null,"abstract":"<div><p>Organ-on-chip (OOC) models can be useful tools for cancer drug discovery. Advances in OOC technology have led to the development of more complex assays, yet analysis of these systems does not always account for these advancements, resulting in technical challenges. A challenging task in the analysis of these two-channel microfluidic models is to define the boundary between the channels so objects moving within and between channels can be quantified. We propose a novel imaging-based application of a thin plate spline method – a generalized cubic spline that can be used to model coordinate transformations – to model a tissue boundary and define compartments for quantification of invaded objects, representing the early steps in cancer metastasis. To evaluate its performance, we applied our analytical approach to an adapted OOC developed by Emulate, Inc., utilizing a two-channel system with endothelial cells in the bottom channel and colorectal cancer (CRC) patient-derived organoids (PDOs) in the top channel. Initial application and visualization of this method revealed boundary variations due to microscope stage tilt and ridge and valley-like contours in the endothelial tissue surface. The method was functionalized into a reproducible analytical process and web tool – the Chip Invasion and Contour Analysis (ChICA) – to model the endothelial surface and quantify invading tumor cells across multiple chips. To illustrate applicability of the analytical method, we applied the tool to CRC organoid-chips seeded with two different endothelial cell types and measured distinct variations in endothelial surfaces and tumor cell invasion dynamics. Since ChICA utilizes only positional data output from imaging software, the method is applicable to and agnostic of the imaging tool and image analysis system used. The novel thin plate spline method developed in ChICA can account for variation introduced in OOC manufacturing or during the experimental workflow, can quickly and accurately measure tumor cell invasion, and can be used to explore biological mechanisms in drug discovery.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 4","pages":"Article 100163"},"PeriodicalIF":3.1,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S247255522400025X/pdfft?md5=1d111c27c601b575d3944e3576f55d04&pid=1-s2.0-S247255522400025X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141138799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a generic methyltransferase enzymatic assay based on an SAH riboswitch","authors":"Ha Pham ,&nbsp;Meera Kumar ,&nbsp;Anibal Ramos Martinez ,&nbsp;Mahbbat Ali ,&nbsp;Robert G. Lowery","doi":"10.1016/j.slasd.2024.100161","DOIUrl":"10.1016/j.slasd.2024.100161","url":null,"abstract":"<div><p>Methylation of proteins and nucleic acids plays a fundamental role in epigenetic regulation, and discovery of methyltransferase (MT) inhibitors is an area of intense activity. Because of the diversity of MTs and their products, assay methods that detect <em>S</em>-adenosylhomocysteine (SAH) – the invariant product of <em>S</em>-adenosylmethionine (SAM)-dependent methylation reactions - offer some advantages over methods that detect specific methylation events. However, direct, homogenous detection of SAH requires a reagent capable of discriminating between SAH and SAM, which differ by a single methyl group. Moreover, MTs are slow enzymes and many have submicromolar affinities for SAM; these properties translate to a need for detection of SAH at low nanomolar concentrations in the presence of excess SAM. To meet these needs, we leveraged the exquisite molecular recognition properties of a naturally occurring SAH-sensing RNA aptamer, or riboswitch. By splitting the riboswitch into two fragments, such that SAH binding induces assembly of a trimeric complex, we engineered sensors that transduce binding of SAH into positive fluorescence polarization (FP) and time resolved Förster resonance energy transfer (TR-FRET) signals. The split riboswitch configuration, called the AptaFluor™ SAH Methyltransferase Assay, allows robust detection of SAH (Z’ &gt; 0.7) at concentrations below 10 nM, with overnight signal stability in the presence of typical MT assay components. The AptaFluor assay tolerates diverse MT substrates, including histones, nucleosomes, DNA and RNA, and we demonstrated its utility as a robust, enzymatic assay method for several methyltransferases with SAM K_m values &lt; 1 µM. The assay was validated for HTS by performing a pilot screen of 1,280 compounds against the SARS-CoV-2 RNA capping enzyme, nsp14. By enabling direct, homogenous detection of SAH at low nanomolar concentrations, the AptaFluor assay provides a universal platform for screening and profiling MTs at physiologically relevant SAM concentrations.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 4","pages":"Article 100161"},"PeriodicalIF":3.1,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000236/pdfft?md5=fc3163f869c64a2c1aec56e9077589d4&pid=1-s2.0-S2472555224000236-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141094784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterisation of high throughput screening outputs for small molecule degrader discovery","authors":"Lillie E. Bell ,&nbsp;Catherine Bardelle ,&nbsp;Martin J Packer ,&nbsp;Johanna Kastl ,&nbsp;Geoffrey A. Holdgate ,&nbsp;Gareth Davies","doi":"10.1016/j.slasd.2024.100162","DOIUrl":"10.1016/j.slasd.2024.100162","url":null,"abstract":"<div><p>Targeted protein degradation is an important mechanism carried out by the cellular machinery, one that is gaining momentum as an exploitable strategy for the development of drug-like compounds. Molecules which are able to induce proximity between elusive therapeutic targets of interest and E3 ligases which subsequently leads to proteasomal degradation of the target are beginning to decrease the percentage of the human proteome described as undruggable. Therefore, having the ability to screen for, and understand the mechanism of, such molecules is becoming an increasingly attractive scientific focus. We have established a number of cascade experiments including cell-based assays and orthogonal triage steps to provide annotation to the selectivity and mechanism of action for compounds identified as putative degraders from a primary high throughput screen against a high value oncology target. We will describe our current position, using PROTACs as proof-of-concept, on the analysis of these novel outputs and highlight challenges encountered.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 5","pages":"Article 100162"},"PeriodicalIF":3.1,"publicationDate":"2024-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000248/pdfft?md5=dc288a87ff98b6fd26cc25724269aadb&pid=1-s2.0-S2472555224000248-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141140527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Creating a more strategic small molecule biophysical hit characterization workflow","authors":"Christopher Fotsch ,&nbsp;Debaleena Basu ,&nbsp;Ryan Case ,&nbsp;Qing Chen ,&nbsp;Pratibha C. Koneru ,&nbsp;Mei-Chu Lo ,&nbsp;Rachel Ngo ,&nbsp;Pooja Sharma ,&nbsp;Amit Vaish ,&nbsp;Xiang Yi ,&nbsp;Stephan G. Zech ,&nbsp;Peter Hodder","doi":"10.1016/j.slasd.2024.100159","DOIUrl":"10.1016/j.slasd.2024.100159","url":null,"abstract":"<div><p>To confirm target engagement of hits from our high-throughput screening efforts, we ran biophysical assays on several hundreds of hits from 15 different high-throughput screening campaigns. Analyzing the biophysical assay results from these screening campaigns led us to conclude that we could be more strategic in our biophysical analysis of hits by first confirming activity in a thermal shift assay (TSA) and then confirming activity in either a surface plasmon resonance (SPR) assay or a temperature-related intensity change (TRIC) assay. To understand how this new workflow shapes the quality of the final hits, we compared TSA/SPR or TSA/TRIC confirmed and unconfirmed hits to one another using four measures of compound quality: quantitative estimate of drug-likeness (QED), Pan-Assay Interference Compounds (PAINS), promiscuity, and aqueous solubility. In general, we found that the biophysically confirmed hits performed better in the compound quality metrics than the unconfirmed hits, demonstrating that our workflow not only confirmed target engagement of the hits but also enriched for higher quality hits.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 4","pages":"Article 100159"},"PeriodicalIF":3.1,"publicationDate":"2024-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000212/pdfft?md5=389ec9b1863fb13b54ef5a3823355f2d&pid=1-s2.0-S2472555224000212-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140900579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing scaffold-free spheroid models: 3D cell bioprinting method for metastatic HSC3–Oral squamous carcinoma cell line","authors":"Taís Bacelar Sacramento de Araújo ,&nbsp;Raphael Luís Rocha Nogueira ,&nbsp;Leonardo de Oliveira Siquara da Rocha ,&nbsp;Iasmin Nogueira Bastos ,&nbsp;Rosane Borges Dias ,&nbsp;Bruno Solano De Freitas Souza ,&nbsp;Daniel William Lambert ,&nbsp;Ricardo D. Coletta ,&nbsp;Viviane Aline Oliveira Silva ,&nbsp;Clarissa A. Gurgel Rocha","doi":"10.1016/j.slasd.2024.100158","DOIUrl":"https://doi.org/10.1016/j.slasd.2024.100158","url":null,"abstract":"<div><p>3D <em>in vitro</em> systems offer advantages over the shortcomings of two-dimensional models by simulating the morphological and functional features of <em>in vivo</em>-like environments, such as cell-cell and cell-extracellular matrix interactions, as well as the co-culture of different cell types. Nevertheless, these systems present technical challenges that limit their potential in cancer research requiring cell line- and culture-dependent standardization. This protocol details the use of a magnetic 3D bioprinting method and other associated techniques (cytotoxicity assay and histological analysis) using oral squamous cell carcinoma cell line, HSC3, which offer advantages compared to existing widely used approaches. This protocol is particularly timely, as it validates magnetic bioprinting as a method for the rapid deployment of 3D cultures as a tool for compound screening and development of heterotypic cultures such as co-culture of oral squamous cell carcinoma cells with cancer-associated fibroblasts (HSC3/CAFs).</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 4","pages":"Article 100158"},"PeriodicalIF":3.1,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000200/pdfft?md5=65af641e1cc59f0ba7520fa27146fec1&pid=1-s2.0-S2472555224000200-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140924481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"POLARISED views and FRETting about probe modulation assays: Learning from High Throughput Screening","authors":"Alice Lanne ,&nbsp;Catherine Bardelle ,&nbsp;Gareth Davies ,&nbsp;Antonia Turberville ,&nbsp;Hannah Semple ,&nbsp;Rachel Moore ,&nbsp;Geoffrey A. Holdgate","doi":"10.1016/j.slasd.2024.100156","DOIUrl":"https://doi.org/10.1016/j.slasd.2024.100156","url":null,"abstract":"<div><p>Fluorescent probe modulation assays are a widely used approach to monitor displacement or stabilisation of fluorescently labelled tool ligands by test compounds. These assays allow an optical read-out of probe-receptor binding and can be used to detect compounds that compete with the labelled ligand, either directly or indirectly. Probes for both orthosteric and allosteric sites are often employed. The method can also be used to identify test compounds that may stabilise the ternary complex, offering an opportunity to discover novel molecular glues. The utility of these fluorescence-based assays within high-throughput screening has been facilitated by the use of streptavidin labelled terbium as a donor and access to a range of different acceptor fluorophores. During 2023, the High-throughput Screening group at AstraZeneca carried out 8 high-throughput screens using these approaches. In this manuscript we will present the types of assays used, an overview of the timelines for assay development and screening, the application of orthogonal artefact methods to aid hit finding and the results of the screens in terms of hit rate and the number of compounds identified with IC₅₀ values of better than 30 µM. Learning across the development, execution and analysis of these screens will be presented.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 4","pages":"Article 100156"},"PeriodicalIF":3.1,"publicationDate":"2024-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000182/pdfft?md5=31508b5b2d14cbf4e0c02d5d7e69cecf&pid=1-s2.0-S2472555224000182-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140645364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aldehyde Dehydrogenase 2 (ALDH2): A novel sorafenib target in hepatocellular carcinoma unraveled by the proteome-wide cellular thermal shift assay","authors":"Inês C. Ferreira ,&nbsp;Estefania Torrejón ,&nbsp;Bernardo Abecasis ,&nbsp;Bruno M. Alexandre ,&nbsp;Ricardo A. Gomes ,&nbsp;Chris Verslype ,&nbsp;Jos van Pelt ,&nbsp;Ana Barbas ,&nbsp;Daniel Simão ,&nbsp;Tiago M. Bandeiras ,&nbsp;Alessio Bortoluzzi ,&nbsp;Sofia P. Rebelo","doi":"10.1016/j.slasd.2024.100154","DOIUrl":"10.1016/j.slasd.2024.100154","url":null,"abstract":"<div><p>Sorafenib is a multikinase inhibitor indicated for first-line treatment of unresectable hepatocellular carcinoma. Despite its widespread use in the clinic, the existing knowledge of sorafenib mode-of-action remains incomplete. To build upon the current understanding, we used the Cellular Thermal Shift Assay (CETSA) coupled to Mass Spectrometry (CETSA-MS) to monitor compound binding to its target proteins in the cellular context on a proteome-wide scale. Among the potential sorafenib targets, we identified aldehyde dehydrogenase 2 (ALDH2), an enzyme that plays a major role in alcohol metabolism. We validated the interaction of sorafenib with ALDH2 by orthogonal methods using pure recombinant protein, proving that this interaction is not mediated by other cellular components. Moreover, we showed that sorafenib inhibits ALDH2 activity, supporting a functional role for this interaction. Finally, we were able to demonstrate that both ALDH2 protein expression and activity were reduced in sorafenib-resistant cells compared to the parental cell line. Overall, our study allowed the identification of ALDH2 as a novel sorafenib target and sheds light on its potential role in both hepatocellular carcinoma and sorafenib resistance condition.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 3","pages":"Article 100154"},"PeriodicalIF":3.1,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000169/pdfft?md5=e39307bdcc31a11896db9724bd824787&pid=1-s2.0-S2472555224000169-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140195277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment, optimization and validation of a fluorescence polarization-based high-throughput screening assay targeting cathepsin L inhibitors","authors":"Wenwen Zhou ,&nbsp;Baoqing You ,&nbsp;Xiaomeng Zhao ,&nbsp;Shuyi Si ,&nbsp;Yan Li ,&nbsp;Jing Zhang","doi":"10.1016/j.slasd.2024.100153","DOIUrl":"10.1016/j.slasd.2024.100153","url":null,"abstract":"<div><p>Cathepsin L (CTSL), a lysosomal cysteine proteinase, is primarily dedicated to the metabolic turnover of intracellular proteins. It is involved in various physiological processes and contributes to pathological conditions such as viral infection, tumor invasion and metastasis, inflammatory status, atherosclerosis, renal disease, diabetes, bone diseases, and other ailments. The coronavirus disease 2019 (COVID-19), with its rapid global spread and significant mortality, has been a worldwide epidemic since the late 2019s. Notably, CTSL plays a role in the processing of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein, providing a potential avenue to block coronavirus host cell entry and thereby inhibit SARS-CoV-2 infection in humans. In this study, we have developed a novel method using fluorescence polarization (FP) for screening CTSL inhibitors in a high-throughput format. The optimized assay demonstrated its appropriateness for high-throughput screening (HTS) with a Z-factor of 0.9 in a 96-well format. Additionally, the IC₅₀ of the known inhibitor, Z-Phe-Tyr-CHO, was determined to be 188.50 ± 46.88 nM. Upon screening over 2000 small molecules, we identified, for the first time, the anti-CTSL properties of a benzothiazoles derivative named IMB 8015. This work presents a novel high-throughput approach and its application in discovering and evaluating CTSL inhibitors.</p></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"29 3","pages":"Article 100153"},"PeriodicalIF":3.1,"publicationDate":"2024-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2472555224000157/pdfft?md5=531dea274a50b4826ec3758fe007d67d&pid=1-s2.0-S2472555224000157-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140190478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}